| 15578574 |
Yamaguchi M, Uchiyama S: Regucalcin stimulates osteoclast-like cell formation in mouse marrow cultures. J Cell Biochem. 2005 Mar 1;94(4):794-803.
The effect of regucalcin, a regulatory protein in intracellular signaling, on osteoclastic cell formation in mouse bone marrow culture is investigated. The bone marrow cells were cultured for 7 days in an alpha-minimal essential medium containing either vehicle or regucalcin (10 (-10)-10 (-8) M). Osteoclast-like cell formation was estimated by staining for tartrate-resistant acid phosphatase (TRACP), a marker enzyme of osteoclasts. The presence of regucalcin (10 (-10)-10 (-8) M) caused a remarkable increase in osteoclast-like multinucleated cells (MNCs). The effect of regucalcin in stimulating osteoclast-like cell formation was significantly inhibited in the presence of calcitonin (CT; 10 (-9) M), 17beta-estradiol (10 (-9) M), beta-cryptoxanthin (CX; 10 (-6) M), or zinc sulfate (10 (-4) M), which is an anti-bone resorbing factor. The effect of regucalcin on osteoclast-like cell formation was not significantly blocked in the presence of cycloheximide, an inhibitor of protein synthesis, or 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB), an inhibitor of transcriptional activity. The effect of parathyroid hormone (10 (-7) M), 1,25-dihydroxyvitamin D (3) (10 (-7) M), prostaglandin E (2) (10 (-5) M), or tumor necrosis factor-alpha (10 ng/ml) in increasing osteoclast-like cell formation was significantly enhanced in the presence of regucalcin (10 (-8) M). Moreover, when rat femoral-diaphyseal or -metaphyseal tissues were cultured for 48 h in the presence of regucalcin (10 (-10)-10 (-8) M), the diaphyseal or metaphyseal calcium content was significantly decreased in the presence of regucalcin (10 (-10)-10 (-8) M) in vitro. The consumption of glucose and the production of lactic acid in culture medium by the diaphyseal or metaphyseal tissues was significantly raised in the presence of regucalcin (10 (-10)-10 (-8) M). This study demonstrates that regucalcin directly stimulates osteoclast-like cell formation in mouse marrow culture in vitro, and that the protein stimulates bone resorption in rat femoral tissues in vitro. |
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